Chapter 19. Control of Eukaryotic Genome. AP Biology
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1 Chapter 19. Control of Eukaryotic Genome
2 The BIG Questions How are genes turned on & off in eukaryotes? How do cells with the same genes differentiate to perform completely different, specialized functions?
3 Prokaryote vs. eukaryote genome Prokaryotes small size of genome circular molecule of naked DNA DNA is readily available to RNA polymerase control of transcription by regulatory proteins operon system most of DNA codes for protein or RNA no introns, small amount of non-coding DNA regulatory sequences: promoters, operators
4 Prokaryote vs. eukaryote genome Eukaryotes much greater size of genome how does all that DNA fit into nucleus? DNA packaged in chromatin fibers regulates access to DNA by RNA polymerase cell specialization need to turn on & off large numbers of genes most of DNA does not code for protein 97% junk DNA in humans
5 Points of control The control of gene expression can occur at any step in the pathway from gene to functional protein unpacking DNA transcription mrna processing mrna transport out of nucleus through cytoplasm protection from degradation translation protein processing protein degradation
6 Why turn genes on & off? Specialization each cell of a multicellular eukaryote expresses only a small fraction of its genes Development different genes needed at different points in life cycle of an organism afterwards need to be turned off permanently Responding to organism s needs homeostasis cells of multicellular organisms must continually turn certain genes on & off in response to signals from their external & internal environment
7 DNA packing How do you fit all that DNA into nucleus? DNA coiling & folding double helix nucleosomes chromatin fiber looped domains chromosome from DNA double helix to condensed chromosome
8 Nucleosomes Beads on a string 1 st level of DNA packing histone proteins 8 protein molecules many positively charged amino acids arginine & lysine bind tightly to negatively charged DNA 8 histone molecules DNA packing movie
9 DNA packing Degree of packing of DNA regulates transcription tightly packed = no transcription = genes turned off darker DNA (H) = tightly packed lighter DNA (E) = loosely packed
10 DNA methylation Methylation of DNA blocks transcription factors no transcription = genes turned off attachment of methyl groups ( CH 3 ) to cytosine C = cytosine nearly permanent inactivation of genes ex. inactivated mammalian X chromosome
11 Histone acetylation Acetylation of histones unwinds DNA loosely packed = transcription = genes turned on attachment of acetyl groups ( COCH 3 ) to histones conformational change in histone proteins transcription factors have easier access to genes
12 Transcription initiation Control regions on DNA promoter nearby control sequence on DNA binding of RNA polymerase & transcription factors base rate of transcription enhancers distant control sequences on DNA binding of activator proteins enhanced rate (high level) of transcription
13 Model for Enhancer action Enhancer DNA sequences distant control sequences Activator proteins bind to enhancer sequence & stimulates transcription Silencer proteins bind to enhancer sequence & block gene transcription Turning on Gene movie
14 Post-transcriptional control Alternative RNA splicing variable processing of exons creates a family of proteins
15 Regulation of mrna degradation Life span of mrna determines pattern of protein synthesis mrna can last from hours to weeks RNA processing movie
16 RNA interference Small RNAs (srna) NEW! short segments of RNA (21-28 bases) bind to mrna create sections of double-stranded mrna death tag for mrna triggers degradation of mrna cause gene silencing even though post-transcriptional control, still turns off a gene sirna
17 RNA interference Small RNAs mrna Hottest new topic in biology double-stranded RNA srna + mrna mrna degraded functionally turns gene off
18 Control of translation Block initiation stage regulatory proteins attach to 5 end of mrna prevent attachment of ribosomal subunits & initiator trna block translation of mrna to protein Control of translation movie
19 Protein processing & degradation Protein processing folding, cleaving, adding sugar groups, targeting for transport Protein degradation ubiquitin tagging proteosome degradation Protein processing movie
20 Ubiquitin 1980s 2004 Death tag mark unwanted proteins with a label 76 amino acid polypeptide, ubiquitin labeled proteins are broken down rapidly in "waste disposers" proteasomes Aaron Ciechanover Israel Avram Hershko Israel Irwin Rose UC Riverside
21 Proteasome Protein-degrading machine cell s waste disposer can breakdown all proteins into 7-9 amino acid fragments play Nobel animation
22 6 posttranslation 1. transcription -DNA packing -transcription factors 2. mrna processing -splicing 1 5 translation 4 mrna transport in cytoplasm 3. mrna transport out of nucleus -breakdown by srna 4. mrna transport in cytoplasm -protection by 3 cap & poly-a tail transcription 5. translation -factors which block start of translation 2 mrna processing 3 mrna transport out of nucleus 6. post-translation -protein processing -protein degradation
23 Any Questions??
24
25 Structure of the Eukaryotic Genome
26 How many genes? Genes only ~3% of human genome protein-coding sequences 1% of human genome non-protein coding genes 2% of human genome trna ribosomal RNAs sirnas
27 What about the rest of the DNA? Non-coding DNA sequences regulatory sequences promoters, enhancers terminators junk DNA introns repetitive DNA centromeres telomeres tandem & interspersed repeats transposons & retrotransposons Alu in humans
28 Repetitive DNA Repetitive DNA & other non-coding sequences account for most of eukaryotic DNA
29 Genetic disorders of repeats Fragile X syndrome most common form of inherited mental retardation defect in X chromosome mutation of FMR1 gene causing many repeats of CGG triplet in promoter region 200+ copies normal = 6-40 CGG repeats FMR1 gene not expressed & protein (FMRP) not produced function of FMR1 protein unknown binds RNA
30 Fragile X syndrome The more triplet repeats there are on the X chromosome, the more severely affected the individual will be mutation causes increased number of repeats (expansion) with each generation
31 Huntington s Disease Rare autosomal dominant degenerative neurological disease 1 st described in 1872 by Dr. Huntington most common in white Europeans 1 st symptoms at age death comes ~12 years after onset Mutation on chromosome 4 CAG repeats copies normal = CAG repeats CAG codes for glutamine amino acid
32 Huntington s disease Abnormal (huntingtin) protein produced chain of charged glutamines in protein bonds tightly to brain protein, HAP-1 Woody Guthrie
33 Families of genes Human globin gene family evolved from duplication of common ancestral globin gene Different versions are expressed at different times in development allowing hemoglobin to function throughout life of developing animal
34 Hemoglobin differential expression of different beta globin genes ensures important physiological changes during human development
35 Interspersed repetitive DNA Repetitive DNA is spread throughout genome interspersed repetitive DNA make up 25-40% of mammalian genome in humans, at least 5% of genome is made of a family of similar sequences called, Alu elements 300 bases long Alu is an example of a "jumping gene" a transposon DNA sequence that "reproduces" by copying itself & inserting into new chromosome locations
36 Rearrangements in the genome Transposons transposable genetic element piece of DNA that can move from one location to another in cell s genome One gene of an insertion sequence codes for transposase, which catalyzes the transposon s movement. The inverted repeats, about 20 to 40 nucleotide pairs long, are backward, upside-down versions of each oth. In transposition, transposase molecules bind to the inverted repeats & catalyze the cutting & resealing of DNA required for insertion of the transposon at a target site.
37 Transposons Insertion of transposon sequence in new position in genome insertion sequences cause mutations when they happen to land within the coding sequence of a gene or within a DNA region that regulates gene expression
38 Transposons Barbara McClintock discovered 1st transposons in Zea mays (corn) in 1947
39
40 Retrotransposons Transposons actually make up over 50% of the corn (maize) genome & 10% of the human genome. Most of these transposons are retrotransposons, transposable elements that move within a genome by means of RNA intermediate, transcript of the retrotransposon DNA
41 Any Questions??
42 Aaaaah Structure-Function yet again!
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1. Please identify the molecule below: 5 -ACTCGATTACGATACGA-3ʼ a) DNA b) mrna c) trna d) rrna e) It cannot be determined 2. If a complimentary strand of RNA were made to the molecule in question 1, what
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